Methods for predicting risk of developing hypertension

ABSTRACT

Methods using biomarkers, e.g., serum levels of ST2, to predict risk of developing hypertension, as well as methods for treating subjects to reduce the risk of developing hypertension and methods for selecting and/or stratifying subjects for clinical trials of treatments to reduce the risk of hypertension.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.13/969,116, filed Aug. 16, 2013 (issued as U.S. Pat. No. 9,523,696),which claims priority to U.S. patent application Ser. No. 61/683,956,filed Aug. 16, 2012, the contents of which are incorporated by referencein their entirety.

TECHNICAL FIELD

This invention relates to the field of molecular biology andcardiovascular medicine, including methods using biomarkers, e.g., serumlevels of ST2, to predict risk of developing hypertension, as well asmethods for treating subjects to reduce the risk of developinghypertension and methods for selecting and/or stratifying subjects forclinical trials of treatments to reduce the risk of hypertension.

BACKGROUND OF THE INVENTION

Hypertension, often referred to colloquially as “high blood pressure” isa condition characterized by the presence of a systolic bloodpressure≥140 mmHg and a diastolic blood pressure≥90 mmHg (referred to as140/90). Blood pressures between 120/80 and 140/90 are typicallyconsidered prehypertension, while pressure below 120/80 is normal. Otherthan pregnancy, the treatment of hypertension is the most common reasonfor physician office visits and use of prescription drugs among USadults (Egan et al., JAMA 303(20):2043, 2010). Hypertension is a majorrisk factor for cardiovascular disease: an estimated 69% of patientswith incident myocardial infarction, and 74% with incident heart failurehave preceding hypertension (Roger et al., Circulation 125:e2-e220,2012). Treatment and control of hypertension reduces the risk of thesecardiovascular diseases (Meredith, Journal ofRenin-Angiotensin-Aldosterone System, 7(2):64-73, 2006). Therefore,identifying subjects who are at risk of developing hypertension, andtreating them to reduce that risk, would reduce their risk ofcardiovascular disease.

SUMMARY

The invention is based, at least in part, on the discovery that subjectshaving an elevated level of soluble ST2 have an increased risk ofdeveloping hypertension. Thus, provided herein are methods for selectinga treatment for a subject that include determining a level of solubleST2 in a biological sample from a subject, comparing the level ofsoluble ST2 in the biological sample to a reference level of solubleST2, and selecting an anti-hypertensive treatment (anti-hypertensivetherapy) for a subject having an elevated level of soluble ST2 in thebiological sample compared to the reference level of soluble ST2. Alsoprovided are methods of treating a subject that include determining alevel of soluble ST2 in a biological sample from a subject, comparingthe level of soluble ST2 in the biological sample to a reference levelof soluble ST2, and administering to a subject having an elevated levelof soluble ST2 in the biological sample compared to the reference levelof soluble ST2 an anti-hypertensive agent. Also provided are methods forselecting a subject for participation in a clinical study of a treatmentfor reducing the risk of developing hypertension, and methods ofevaluating the risk of developing hypertension in a subject that includedetermining a level of soluble ST2 in a biological sample from asubject. Also provided are kits that contain an antibody thatspecifically binds to soluble ST2 and instructions for performing any ofthe methods described herein.

Also provided is the use of a pharmaceutical agent for reducinghypertension (e.g., any of exemplary pharmaceutical agents for reducinghypertension described herein) for treating a subject identified ashaving an increased risk of developing hypertension (using any of themethods described herein).

By the term hypertension is meant a medical condition that ischaracterized by an abnormal, elevated blood pressure (i.e., a systolicblood pressure≥140 mmHg, a diastolic blood pressure≥90 mmHg).

By the term “soluble ST2” is meant a soluble protein containing asequence at least 90% identical (e.g., at least 95%, 96%, 97%, 98%, 99%,or 100% identical) to NCBI Accession No. NP_003847.2 (SEQ ID NO: 1) orcontaining a sequence at least 90% identical (e.g., at least 95%, 96%,97%, 98%, 99%, or 100% identical) to amino acids 19-328 of SEQ ID NO: 1,or a nucleic acid containing a sequence at least 90% identical (e.g., atleast 95%, 96%, 97%, 98%, 99%, or 100% identical) to NCBI Accession No.NM_003856.2 or containing a sequence at least 90% identical (e.g., atleast 95%, 96%, 97%, 98%, 99%, or 100% identical) to nucleotides 285 to1214 of NCBI Accession No. NM_003856.2.

By the term “elevated” or “elevation” is meant a difference, e.g., astatistically significant or detectable increase in a determined ormeasured level (e.g., a human soluble ST2 protein level) compared to areference level (e.g., a level of human soluble ST2 in a subject (orpopulation of subjects) not having an increased risk of developinghypertension, or a threshold level of human soluble ST2). In someembodiments, the reference is a threshold level, and any level abovethat is considered “elevated.” Additional reference levels of humansoluble ST2 are described herein and are known in the art.

As used herein, a “biological sample” includes one or more of blood,serum, plasma, urine, and body tissue. Generally, a biological sample isa sample containing serum, blood, or plasma.

By the term “health care facility” is meant a location where a subjectmay receive medical care or treatment from a health care professional(e.g., a nurse, a physician, or a physician's assistant). Non-limitingexamples of health care facilities include hospitals, clinics, surgicalcenters, and assisted care facilities (e.g., a nursing home).

By the term “reference level” is meant a threshold level or a level in acontrol subject or control patient population. A reference level willdepend on the assay performed and can be determined by one of ordinaryskill in the art. A reference level may be a baseline level or a levelin the same patient measured at an earlier point in time. In someembodiments, a reference level is a level of soluble ST2 in a controlsubject or population of control subjects that does not have anincreased risk of developing hypertension. In some embodiments, areference level is a level of soluble ST2 in a healthy subject.Additional examples of reference levels of soluble ST2 and methods ofdetermining the same are known in the art and are described herein.

In some embodiments, the ratio of two soluble ST2 levels in a subject iscompared to a reference ratio (e.g., a ratio of soluble ST2 levelsmeasured in a control subject, e.g., any of the control subjectsdescribed herein or the same subject at earlier time points). Additionalexamples of reference ratios of soluble ST2 are known in the art and aredescribed herein.

As used herein, a “subject” is a mammal, e.g., a human. In allembodiments, human nucleic acids, human polypeptides, and human subjectscan be used.

By the term “healthy subject” is meant a subject that does not have adisease (e.g., a cardiac disease). For example, a healthy subject hasnot been diagnosed as having a disease and is not presenting with two ormore (e.g., two, three, four, or five) symptoms of a disease state.

By the term “disease state” is meant the manifestation of one or more(e.g., at least two, three, four, or five) symptoms in a subject thatindicate either an abnormal decrease in the viability and/or an abnormaldecrease/malfunction of a biological activity of one or more (e.g., atleast two, three, four, or five) tissues in the body of the subject.Non-limiting examples of disease states in a subject include a cardiacdisease (e.g., arrhythmia, heart failure, heart attack, coronary arterydisease, cardiovascular disease, acute coronary syndrome, and angina),inflammation, stroke, renal failure, obesity, high cholesterol, anddyslipidemia.

By the phrase “physical symptoms associated with a disease state” ismeant the one or more (e.g., at least two, three, or four) symptoms thatare manifested by a subject having a particular disease state. Physicalsymptoms associated with several disease states are known in the art bymedical health professionals (e.g., physicians). Non-limiting examplesof physical symptoms associated with a cardiac disease (e.g.,arrhythmia, heart failure, coronary artery disease, cardiovasculardisease, acute coronary syndrome, and angina) include shortness ofbreath, heart palpitations, increased heart rate, weakness, dizziness,nausea, sweating, chest discomfort or pressure, chest pain, arm pain,fullness, indigestion, sweating, wheezing, sleep apnea, and anxiety.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Methods and materials aredescribed herein for use in the present invention; other, suitablemethods and materials known in the art can also be used. The materials,methods, and examples are illustrative only and not intended to belimiting. All publications, patent applications, patents, sequences,database entries, and other references mentioned herein are incorporatedby reference in their entirety. In case of conflict, the presentspecification, including definitions, will control.

The details of one or more embodiments of the invention are set forth inthe accompanying drawings and the description below. Other features,objects, and advantages of the invention will be apparent from thedescription and drawings, and from the claims.

DETAILED DESCRIPTION

The invention is based, in part, on the discovery that subjects havingan elevated level of soluble ST2 or an increase in soluble ST2 over timehave an increased risk of developing hypertension. Thus, provided hereinare methods for identifying subjects who are at increased risk ofdeveloping hypertension. Also described herein are methods for selectinga treatment for a subject that include determining a level of solubleST2 in a biological sample from a subject, comparing the level ofsoluble ST2 in the biological sample to a reference level of solubleST2, and selecting an anti-hypertensive therapy (anti-hypertensivetreatment) for a subject having an elevated level of soluble ST2 in thebiological sample compared to the reference level of soluble ST2. Alsoprovided are methods of treating a subject that include determining alevel of soluble ST2 in a biological sample from a subject, comparingthe level of soluble ST2 in the biological sample to a reference levelof soluble ST2, and administering to a subject having an elevated levelof soluble ST2 in the biological sample compared to the reference levelof soluble ST2 an anti-hypertensive therapy (anti-hypertensivetreatment). Also provided are methods for selecting a subject forparticipation in a clinical study of a treatment for reducing the riskdeveloping hypertension, and methods of evaluating the risk ofdeveloping hypertension in a subject that include determining a level ofsoluble ST2 in a biological sample from a subject. Also provided arekits that contain an antibody that specifically binds to soluble ST2 andinstructions for performing any of the methods described herein.

ST2

The ST2 gene is a member of the interleukin-1 receptor family, whoseprotein product exists both as a trans-membrane form, as well as asoluble receptor that is detectable in serum (Kieser et al., FEBS Lett.372(2-3):189-93 (1995); Kumar et al., J. Biol. Chem. 270(46):27905-13(1995); Yanagisawa et al., FEBS Lett. 302(1):51-3 (1992); Kuroiwa etal., Hybridoma 19(2):151-9 (2000)). ST2 was recently described to bemarkedly up-regulated in an experimental model of heart failure(Weinberg et al., Circulation 106(23):2961-6 (2002)), and preliminaryresults suggest that ST2 concentrations may be elevated in those withchronic severe heart failure (Weinberg et al., Circulation 107(5):721-6(2003)), as well in subjects with acute myocardial infarction (MI)(Shimpo et al., Circulation 109(18):2186-90 (2004)).

The transmembrane form of ST2 is thought to play a role in modulatingresponses of T-helper type 2 cells (Lohning et al., Proc. Natl. Acad.Sci. U.S.A. 95(12):6930-6935 (1998); Schmitz et al., Immunity23(5):479-90 (2005)), and may play a role in development of tolerance instates of severe or chronic inflammation (Brint et al., Nat. Immunol.5(4):373-9 (2004)), while the soluble form of ST2 is up-regulated ingrowth stimulated fibroblasts (Yanagisawa et al., 1992, supra).Experimental data suggest that the ST2 gene is markedly up-regulated instates of myocyte stretch (Weinberg et al., 2002, supra) in a manneranalogous to the induction of the BNP gene (Bruneau et al., Cardiovasc.Res. 28(10):1519-25 (1994)).

Tominaga, FEBS Lett. 258:301-304 (1989), isolated murine genes that werespecifically expressed by growth stimulation in BALB/c-3T3 cells; theytermed one of these genes St2. The St2 gene encodes two proteinproducts: ST2 (IL1RL1), which is a soluble secreted form; and ST2L, atransmembrane receptor form that is very similar to the interleukin-1receptors. The HUGO Nomenclature Committee designated the human homologof ST2, the cloning of which was described in Tominaga et al., Biochim.Biophys. Acta. 1171:215-218 (1992), as Interleukin 1 Receptor-Like 1(IL1RL1). The two terms are used interchangeably herein.

The cDNA sequence of the shorter, soluble isoform of human ST2 can befound at GenBank Acc. No. NM_003856.2 (SEQ ID NO: 2), and thepolypeptide sequence is at GenBank Acc. No. NP_003847.2 (SEQ ID NO: 1;shown below). The mRNA sequence for the longer form of human ST2 is atGenBank Acc. No. NM_016232.4; and the polypeptide sequence is at GenBankAcc. No. NP_057316.3. Additional information is available in the publicdatabases at GeneID: 9173, MIM ID #601203, and UniGene No. Hs.66. Ingeneral, in the methods described herein, the soluble form of ST2polypeptide is measured. Non-limiting examples of soluble ST2 proteininclude proteins containing a sequence at least 90% identical (e.g., atleast 95%, 96%, 97%, 98%, 99%, or 100% identical) to the sequence of SEQID NO: 1. Non-limiting examples of soluble ST2 nucleic acids includenucleic acids containing a sequence at least 90% identical (e.g., atleast 95%, 96%, 97%, 98%, 99%, or 100% identical) to the sequence ofNCBI Accession No. NM_003856.2 (SEQ ID NO: 2).

Human Soluble ST2 Protein (SEQ ID NO: 1)   1mgfwilailt ilmystaakf skqswglene alivrcprqg kpsytvdwyy sqtnksipt  61ernrvfasgq llkflpaava dsgiytcivr sptfnrtgya nvtiykkqsd cnvpdylmys 121tvsgseknsk iycptidlyn wtaplewfkn cqalqgsryr ahksflvidn vmtedagdyt 181ckfihnenga nysvtatrsf tvkdeqgfsl fpvigapaqn eikeveigkn anltcsacfg 241kgtqflaavl wqlngtkitd fgepriqqee gqnqsfsngl acldmvlria dvkeedlllq 301ydclalnlhg lrrhtvrlsr knpskecf

Methods for detecting and measuring soluble ST2 are known in the art,e.g., as described in U.S. Patent Application Publication Nos.2003/0124624, 2004/0048286, and 2005/0130136, the entire contents ofeach of which are incorporated herein by reference. In some embodiments,the methods include determining the identity of the nucleotide sequenceat RefSNP ID: rs1041973.

Kits for measuring soluble ST2 polypeptide are also commerciallyavailable, e.g., the ST2 ELISA Kit manufactured by Medical & BiologicalLaboratories Co., Ltd. (MBL International Corp., Woburn, Mass.), No.7638, and the Presage® ST2 Assay, Critical Care Diagnostics, San Diego,Calif. In addition, devices for measuring soluble ST2 and otherbiomarkers are described in U.S. Patent Publication No. 2005/0250156.Levels of soluble ST2 protein can also be measured using the antibodiesproduced from the hybridoma deposited at American Type CultureCollection and designated by Patent Deposit Designation PTA-10432, andthose antibodies described in U.S. Patent Application Publication No.2011/0256635 and WO 2011/127412 (each of which is herein incorporated byreference).

ST2 Reference Levels

Once a level of soluble ST2 has been determined in a biological samplefrom a subject, the level may be compared to a reference level (e.g.,any of the reference levels described herein or known in the art). Insome embodiments, e.g., where the level of soluble ST2 is determinedusing an ELISA, the reference level may represent a threshold level,above which the subject is identified as having an increased risk ofdeveloping hypertension or selected for participation in a clinicalstudy of a treatment for preventing or reducing the risk of developinghypertension. The reference level chosen may depend on the methodology(e.g., the particular antibody or ELISA kit) used to measure the levelsof soluble ST2. Reference levels of soluble ST2 are known in the art andmay readily be determined by one skilled in the art. In someembodiments, the threshold or reference level is the median level of ST2in a population of subjects, wherein those above the median level havean increased risk of developing hypertension. In some embodiments, thethreshold or reference level is a level that represents a cut-off whenthe population is divided, e.g., divided by risk or by ST2 levels, e.g.,the cut-off level for the top quartile or the top tertile.

Non-limiting threshold levels of soluble ST2 may represent the medianlevel of soluble ST2 in particular patient populations, e.g., subjectswith an increased risk of developing hypertension; in some embodiments,the subjects are stratified by various characteristics, e.g., having aBMI of less than 25, 25-29, or over 30; subjects with normal or impairedrenal function; subjects without a cardiac disease (e.g., without adiagnosis of any of the cardiac diseases described herein); or healthy(e.g., undiagnosed with disease, having a low risk of developingdisease, and not presenting with two or more symptoms of a disease) men,women, or children. For example, a threshold value for soluble ST2 mayfall within the range of about 1.0 to 10 ng/mL, 5.0 ng/mL to 10 ng/mL,about 10.0 ng/mL to 20.0 ng/mL, about 10.0 ng/mL to 15.0 ng/mL, about15.0 ng/mL to 20.0 ng/mL, about 20.0 ng/ml to 40 ng/mL, about 20 ng/mLto 30 ng/mL, about 20 ng/mL to 25 ng/mL, about 25 ng/mL to 30 ng/mL,about 30 ng/mL to about 40 ng/mL, about 30 ng/mL to 35 ng/mL, about 35ng/mL to 40 ng/mL, about 40 ng/mL to about 60 ng/mL, about 40 ng/mL toabout 50 ng/mL, and about 50 ng/mL to about 60 ng/mL. In someembodiments, the threshold value is about 25 ng/ml, e.g., about 24.8ng/ml.

In some embodiments, the threshold value for soluble ST2 in men andwomen may be any value listed in the Table 1. For example, the thresholdvalue of soluble ST2 in men may be between 17.0 ng/mL to 19.0 ng/mL,19.0 ng/mL to 21.0 ng/mL, 21.0 ng/mL to 23.0 ng/mL, 23.0 ng/mL to 25.0ng/mL, 25.0 ng/mL to 27.0 ng/mL, 27.0 ng/mL to 29.0 ng/mL, 29.0 ng/mL to31.0 ng/mL, 31.0 ng/mL to 33.0 ng/mL, 33.0 ng/mL to 35.0 ng/mL, 35.0ng/mL to 37.0 ng/mL, 37.0 ng/mL to 39.0 ng/mL, 39.0 ng/mL to 41.0 ng/mL,41.0 ng/mL to 43.0 ng/mL, 43.0 ng/mL to 45.0 ng/mL, 45.0 ng/mL to 47.0ng/mL, 47.0 ng/mL to 49.0 ng/mL, and 49.0 ng/mL to 51.0 ng/mL. Exemplarythreshold values of soluble ST2 in women may be 12.0 ng/mL to 14.0ng/mL, 14.0 ng/mL to 16.0 ng/mL, 16.0 ng/mL to 18.0 ng/mL, 18.0 ng/mL to20.0 ng/mL, 20.0 ng/mL to 22.0 ng/mL, 22.0 ng/mL to 24.0 ng/mL, 24.0ng/mL to 26.0 ng/mL, 26.0 ng/mL to 28.0 ng/mL, 28.0 ng/mL to 30.0 ng/mL,30.0 ng/mL to 32.0 ng/mL, 32.0 ng/mL to 34.0 ng/mL, 34.0 ng/mL to 36.0ng/mL, 36.0 ng/mL to 38.0 ng/mL, and 38.0 ng/mL to 40.0 ng/mL.

As noted above, a threshold level of soluble ST2 may vary depending onthe methodology used to measure the levels of soluble ST2. For example,if an antibody produced from the hybridoma deposited at American TypeCulture Collection, designated with Patent Deposit Deposition PTA-10432,is used to determine a soluble ST2 level, non-limiting threshold valuesof soluble ST2 may include: below 20 ng/mL, 5 ng/mL to 15 ng/mL, 5.0ng/mL to 10 ng/mL, 10 ng/mL to 20 ng/mL, 10 ng/mL to 15 ng/mL, 14.5ng/mL to 25.3 ng/mL, 15 ng/mL to 25 ng/mL, 15 ng/mL to 20 ng/mL, 18.0ng/mL to 20.0 ng/mL, 18.1 ng/mL to 19.9 ng/mL, 20 ng/mL to 30 ng/mL, 20ng/mL to 25 ng/mL, 25 ng/mL to 35 ng/mL, 25 ng/mL to 30 ng/mL, 30 ng/mLto 40 ng/mL, 30 ng/mL to 35 ng/mL, 35 ng/mL to 45 ng/mL, 35 ng/mL to 40ng/ml, and 40 ng/mL to 45 ng/mL. Additional soluble ST2 reference valuesthat may be used, when the antibody produced from the hybridomadesignated PTA-10432 is used to determine a soluble ST2 level, include:for women, 12.4 ng/mL to 19.9 ng/mL, 12.0 ng/mL to 20 ng/mL, 15.3 ng/mLto 17.4 ng/mL, 15.0 to 17.0 ng/mL, below 20 ng/mL, and below 18 ng/mL;and for men, less than 31.0 ng/mL, less than 26.0 ng/mL, 17.6 ng/mL to30.6 ng/mL, 17.0 ng/mL to 30.0 ng/mL, 21.3 ng/mL to 25.1 ng/mL, and 21.0ng/mL to 25.0 ng/mL. Additional non-limiting threshold values that maybe used, when a soluble ST2 level is measured using the antibodyproduced from the hybridoma designated PTA-10432, include: 10 ng/mL, 11ng/mL, 12 ng/mL, 13 ng/mL, 14 ng/mL, 15 ng/mL, 16 ng/mL, 17 ng/mL, 18ng/mL, 19 ng/mL, 20 ng/mL, 21 ng/mL, 22 ng/mL, 23 ng/mL, 24 ng/mL, 25ng/mL, 26 ng/mL, 27 ng/mL, 28 ng/mL, 29 ng/mL, 30 ng/mL, or 31 ng/mL.

TABLE 1 Serum ST2 Concentrations in Healthy Males and Females ST2(ng/mL) Percentiles Combined Male Female 2.5 8.0 8.6 7.3 25 14.5 17.612.4 50 18.8 23.6 16.2 75 25.3 30.6 19.9 90 34.3 37.2 23.7 95 37.9 45.429.0 97.5 45.6 48.5 33.1 99 50.2 52.7 39.9

In additional non-limiting examples, when a soluble ST2 level ismeasured using the ST2 ELISA Kit (MBL International Corp., Woburn,Mass.), the threshold levels of soluble ST2 include: 0.1 ng/mL to 0.6ng/mL, 0.2 ng/mL to 0.6 ng/mL, 0.2 ng/mL to 0.5 ng/mL, 0.3 ng/mL to 0.5ng/mL, 0.2 ng/mL to 0.3 ng/mL, 0.3 ng/mL to 0.4 ng/mL, and 0.4 ng/mL to0.5 ng/mL. Additional non-limiting threshold values that may be used,when the ST2 ELISA Kit (MBL International Corp.) is used to measure asoluble ST2 level, include: 0.17 ng/mL, 0.18 ng/mL, 0.19 ng/mL, 0.20ng/mL, 0.21 ng/mL, 0.22 ng/mL, 0.23 ng/mL, 0.24 ng/mL, 0.25 ng/mL, 0.26ng/mL, 0.27 ng/mL, 0.28 ng/mL, or 0.29 ng/mL of blood, serum, or plasma.

In some embodiments, the reference level of soluble ST2 is a level ofsoluble ST2 present in a control subject who does not have and is not atrisk of developing hypertension, as determined by standard empiricalmethods. In some embodiments, the control subject has not been diagnosedas having hypertension, is not at risk of developing hypertension, has abody mass index of less than 25, and has a cholesterol (totalcholesterol, high density lipoprotein, and/or low density lipoprotein),and triglyceride profile within a normal range.

Additional Markers

Some embodiments of all of the methods described herein may furtherinclude determining the level of one or more (e.g., at least two, three,four, or four) additional markers in a biological sample from thesubject. The additional markers may be selected from the group of:proANP, NT-proANP, ANP, proBNP, NT-proBNP, BNP, troponin, CRP,creatinine, Blood Urea Nitrogen (BUN), galectin, liver function enzymes,albumin, and bacterial endotoxin. The one or more additional markers canbe measured in any of the biological samples herein. The presence of anelevated level (e.g., at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 110%, 120%,130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 210%, 220%, 230%, 240%,250%, 260%, 270%, 280%, 290%, or 300%) of one or more (e.g., at leasttwo, three, or four) of proANP, NT-proANP, ANP, proBNP, NT-proBNP, BNP,troponin, CRP, creatinine, Blood Urea Nitrogen (BUN), galectin, liverfunction enzymes, albumin, and bacterial endotoxin in a subject comparedto a reference level for each of these additional biomarkers may furtherindicate that the subject has an increased risk developing hypertension,the subject should receive treatment (e.g., treatment on an inpatientbasis), or the subject should be selected for participation in aclinical study of a treatment to decrease the risk of developinghypertension.

Once a level of an additional biomarker has been determined in abiological sample from a subject, the level may be compared to areference level of the additional biomarker (e.g., any of the referencelevels described herein or known in the art). In some embodiments, e.g.,where the level of an additional biomarker is determined using an ELISA,the reference level may represent a threshold level, above which thesubject is identified as having an increased risk of developinghypertension, selected for anti-hypertensive treatment(anti-hypertensive therapy), or selected for participation in a clinicalstudy of a treatment for preventing development of hypertension. Thereference level of the additional biomarker chosen may depend on themethodology (e.g., the particular antibody or ELISA kit) used to measurethe levels of the additional biomarker. Reference levels of additionalbiomarkers are known in the art and may readily be determined by oneskilled in the art.

Non-limiting threshold levels of additional biomarkers may represent themedian level of an additional biomarker in particular patientpopulations, e.g., subjects with a BMI of less than 25, subjects withnormal renal function, subjects without cardiac disease (e.g., any ofthe cardiac diseases described herein), healthy (e.g., undiagnosed withdisease, having a low risk of developing disease, and not presentingwith two or more symptoms of a disease) men, healthy (e.g., undiagnosedwith disease, having a low risk of developing disease, and notpresenting with two or more symptoms of a disease) women, and healthy(e.g., undiagnosed with disease, having a low risk of developingdisease, and not presenting with two or more symptoms of disease)children.

In some embodiments, the reference level of an additional biomarker is alevel of an additional biomarker present in a healthy subject (e.g., asubject that does not have a disease (e.g., any of the cardiac diseasesdescribed herein), has not been diagnosed as having a disease, and/or isnot presenting with two or more (e.g., two, three, four, or five)symptoms of a disease state). In some embodiments, a reference level ofan additional biomarker is a level of the additional biomarker from thesame subject at an earlier point in time. In some embodiments, thereference level of an additional biomarker is a level of the additionalbiomarker from a subject that does not have a cardiac disease, has notbeen diagnosed as having a cardiac disease, and/or does not have two ormore symptoms associated with a cardiac disease (e.g., arrhythmia, heartfailure, heart attack, coronary artery disease, cardiovascular disease,acute coronary syndrome, and angina), inflammation, stroke, renalfailure, obesity, high cholesterol, or dyslipidemia. In someembodiments, the reference level of an additional biomarker is a levelof the additional biomarker from a subject that has not been diagnosedas having a cardiac disease and is not at risk for developing a cardiacdisease (e.g., arrhythmia, heart failure, heart attack, coronary arterydisease, cardiovascular disease, acute coronary syndrome, and angina).

In some embodiments, the ratio of two different levels of an additionalbiomarker in a subject is compared to a reference ratio of theadditional biomarker. In some embodiments, the reference ratio of anadditional biomarker can be a threshold ratio (e.g., a reference ratioof 1.00, 1.00, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, or 2.0). In someembodiments, the reference ratio of an additional biomarker is a ratioof two levels of the additional biomarker measured in a control subject(e.g., any of the control subjects described herein or the samesubject). For example, a reference ratio of an additional biomarker canbe a ratio of the levels of an additional biomarker collected at twodifferent time points in a healthy subject (e.g., a subject that doesnot have a disease (e.g., any of the cardiac diseases described herein),has not been diagnosed as having a disease, and/or is not presentingwith two or more (e.g., two, three, four, or five) symptoms of a diseasestate). In some embodiments, a reference ratio is a ratio of levels ofan additional biomarker from the same subject at an earlier point intime. In some embodiments, the reference ratio of an additionalbiomarker is a ratio of the levels of an additional biomarker from asubject that does not have a cardiac disease, has not been diagnosed ashaving a cardiac disease, and/or does not have two or more symptomsassociated with a cardiac disease (e.g., arrhythmia, heart failure,heart attack, coronary artery disease, cardiovascular disease, acutecoronary syndrome, and angina). In some embodiments, the reference ratiois a ratio of levels of an additional biomarker from a subject that hasnot been diagnosed as having a cardiac disease and is not at risk fordeveloping a cardiac disease (e.g., arrhythmia, heart failure, heartattack, coronary artery disease, cardiovascular disease, acute coronarysyndrome, and angina).

Methods for determining the levels of these additional markers are knownin the art. Kits for determining these additional markers arecommercially available.

Methods for Selecting a Treatment for a Subject

Provided herein are methods of selecting a treatment for a subject thatinclude determining a level of soluble ST2 in a biological sample from asubject, comparing the level of soluble ST2 in the biological sample toa reference level of soluble ST2 (e.g., any of the reference levels ofsoluble ST2 described herein), and selecting an anti-hypertensivetreatment (anti-hypertensive therapy) (e.g., with an anti-hypertensiveagent, e.g., one or more of Diuretics, Beta-blockers,Angiotensin-converting enzyme inhibitors (ACE inhibitors), AngiotensinII receptor blockers (ARBs), Calcium channel blockers, Alpha-blockers,Centrally acting drugs, Vasodilators, and/or Renin inhibitors; see,e.g., Kaplan, “Systemic hypertension: Treatment.” In: Bonow et al., eds.Braunwald's Heart Disease: A Textbook of Cardiovascular Medicine. 9thed. Philadelphia, Pa.: Saunders Elsevier; 2011:chap 46, which isincorporated herein by reference in its entirety), or increasedmonitoring (e.g., increased frequency of cardiac function tests) for asubject having an elevated level of soluble ST2 in the biological samplecompared to the reference level of soluble ST2, or selecting lowfrequency outpatient monitoring for a subject having a decreased or nosignificant change in the level of soluble ST2 as compared to thereference level of soluble ST2.

Also provided are methods of selecting a treatment for a subject thatinclude determining a first level of soluble ST2 in a biological samplefrom a subject at a first time point, determining a second level ofsoluble ST2 in a biological sample from the subject at a second timepoint, comparing the second level of soluble ST2 to the first level ofsoluble ST2, and selecting an anti-hypertensive treatment (e.g., with ananti-hypertensive agent), e.g., one or more of Diuretics, Beta-blockers,Angiotensin-converting enzyme inhibitors (ACE inhibitors), AngiotensinII receptor blockers (ARBs), Calcium channel blockers, Alpha-blockers,Centrally acting drugs, Vasodilators, and/or Renin inhibitors; see,e.g., Kaplan, “Systemic hypertension: Treatment.” In: Bonow et al., eds.Braunwald's Heart Disease: A Textbook of Cardiovascular Medicine. 9thed. Philadelphia, Pa.: Saunders Elsevier; 2011:chap 46, which isincorporated herein by reference in its entirety) or increasedmonitoring (e.g., increased frequency of cardiac function tests) for asubject having an elevated second level of soluble ST2 compared to thefirst level of soluble ST2, or selecting low frequency outpatientmonitoring for a subject having a decreased or no significant change insecond level of soluble ST2 as compared to the first level of solubleST2.

The methods described herein can be performed by a medical professional(e.g., a physician, a physician's assistant, a nurse, a nurse'sassistant, or a laboratory technician) or veterinary professional. Thesemethods can be performed in a hospital, a clinic, a primary carefacility (e.g., a nursing home), or a clinical laboratory, or anycombination thereof.

In some embodiments, the biological sample, the first biological sample,and/or the second biological sample contain blood, serum, or plasma. Insome embodiments, the biological sample, the first biological sample,and/or the second biological sample are stored (e.g., at a temperaturebelow 25° C., e.g., at a temperature below 15° C. or 0° C.) for a periodof time (e.g., at least 2, 4, 6, 8, 10, 12, 24, 36, or 48 hours) priorto determining the level of soluble ST2 and/or determining the level ofone or more additional biomarkers (e.g., BNP, proBNP, or NT-proBNP).

In some embodiments, the level of soluble ST2 is determined using anenzyme-linked immunosorbent assay (ELISA) (e.g., using any of thesoluble ST2 ELISA kits described herein or known in the art.

Some embodiments further include detecting a level of one or moreadditional biomarkers (e.g., any of the additional biomarkers describedherein, e.g., BNP, proBNP, and NT-proBNP) in the biological sample fromthe subject. In these embodiments, an anti-hypertensive treatment(anti-hypertensive therapy) or increased cardiac monitoring is selectedfor a subject having an elevation in the level of the one or moreadditional biomarkers in the biological sample compared to a referencelevel of the one or more additional biomarkers, or low frequencyoutpatient monitoring is selected for a subject having a decreased or nosignificant change in the level of the one or more additional markers ascompared to the reference level of the one or more additionalbiomarkers. Some embodiments further include determining a first levelof one or more additional biomarkers (e.g., any of the additionalbiomarkers described herein, e.g., BNP, proBNP, and NT-proBNP) in abiological sample from the subject at a first time point, determining asecond level of the one or more additional biomarkers in a biologicalsample at a second time point, comparing the first level and the secondlevel of the one or more additional biomarkers, and selecting ananti-hypertensive treatment (anti-hypertensive therapy) or increasedcardiac monitoring for a subject having an elevated second level of theone or more additional biomarkers compared to the first level of the oneor more additional biomarkers, or selecting low frequency outpatientmonitoring for a subject having a decreased or no significant change inthe second level of the one or more additional markers compared to thefirst level of the one or more additional markers.

Methods of Treating a Subject

Also provided are methods of treating a subject that include determininga level of soluble ST2 in a biological sample from a subject, comparingthe level of soluble ST2 in the biological sample to a reference levelof soluble ST2, identifying the subject as having an increased risk ofdeveloping hypertension (based on an elevated level of soluble ST2compared to the reference level of soluble ST2), and administering tothe subject having an elevated level of soluble ST2 an anti-hypertensiveagent or performing increased cardiac monitoring of the subject, ormonitoring (e.g., monitoring with a low frequency on an outpatientbasis) a subject having a low level of soluble ST2 in the biologicalsample compared to the reference level of soluble ST2.

Also provided are methods of treating a subject that include determininga first level of soluble ST2 in a biological sample from a subject at afirst time point, determining a second level of soluble ST2 in abiological sample from the subject at a second time point, comparing thesecond level to the first level of soluble ST2, and (1) identifying asubject having an elevated second level of soluble ST2 as compared tothe first level of soluble ST2 as having an increased risk of developinghypertension and administering to the subject identified as having anincreased risk of hypertension (based on the comparison of the secondand first soluble ST2 levels) an anti-hypertensive agent or performingincreased cardiac monitoring on the subject, or (2) identifying asubject having a reduced or no significant change in the second level ofsoluble ST2 compared to the first level of soluble ST2 as having areduced risk of developing hypertension and monitoring (e.g., monitoringwith low frequency on an outpatient basis) the subject identified ashaving a reduced risk of developing hypertension (based on thecomparison of the second and first soluble ST2 levels).

Some embodiments further include detecting a level of one or moreadditional biomarkers (e.g., any of the additional biomarkers describedherein, e.g., BNP, proBNP, and NT-proBNP) in a biological sample fromthe subject (e.g., the biological sample, the first biological sample,and/or the second biological sample). In these embodiments, a subjecthaving an elevation in the level of the one or more additionalbiomarkers in the biological sample compared to a reference level of theone or more additional biomarkers (e.g., any of the reference levels ofthe one or more additional biomarkers described herein) is administeredan anti-hypertensive agent or increased cardiac monitoring, or a subjecthaving decrease or no significant change in the level of the one or moreadditional biomarkers compared to a reference level of the one or moreadditional biomarkers is monitored with low frequency on an outpatientbasis. Some embodiments further include determining a first level of oneor more additional biomarkers (e.g., any of the additional biomarkersdescribed herein, e.g., BNP, proBNP, and NT-proBNP) in a biologicalsample from the subject at the first time point, determining a secondlevel of the one or more additional biomarkers in a biological sample atthe second time point, comparing the first level and the second level ofthe one or more additional biomarkers, and administering ananti-hypertensive agent or performing increased cardiac monitoring on asubject having an elevation in the second level of the one or moreadditional biomarkers compared to the first level of the one or moreadditional biomarkers, or monitoring (e.g., monitoring with lowfrequency on an outpatient basis) a subject having a decreased or nosignificant change in the second level of the one or more additionalbiomarkers as compared to the first level of the one or more additionalbiomarkers.

Some embodiments further include administering to the subject one ormore (e.g., two, three, or four) pharmaceutical agents selected from thegroup of: Diuretics, Beta-blockers, Angiotensin-converting enzymeinhibitors (ACE inhibitors), Angiotensin II receptor blockers (ARBs),Calcium channel blockers, Alpha-blockers, Centrally acting drugs,Vasodilators, and/or Renin inhibitors; see, e.g., Kaplan, “Systemichypertension: Treatment.” In: Bonow et al., eds. Braunwald's HeartDisease: A Textbook of Cardiovascular Medicine. 9th ed. Philadelphia,Pa.: Saunders Elsevier; 2011:chap 46, which is incorporated herein byreference in its entirety.

Methods for Determining the Risk of Developing Hypertension

Also provided are methods of evaluating the risk of developinghypertension in a subject that include determining a level of solubleST2 in a biological sample from a subject, comparing the level ofsoluble ST2 in the biological sample to a reference level of solubleST2, and identifying a subject having an elevated level of soluble ST2in the biological sample compared to the reference level of soluble ST2as having an increased risk of developing hypertension, or identifying asubject having no significant change or a decreased level of soluble ST2in the biological sample compared to the reference level of soluble ST2as having a decreased risk of developing hypertension. In someembodiments, a subject having an elevated level of soluble ST2 (relativeto a reference level of soluble ST2) has an increased risk (e.g., anincreased risk of at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%,55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 110%, 120%, 130%,140%, 150%, 160%, 170%, 180%, 190%, or 200% higher than a referencesubject, e.g., a control subject as described herein) of developinghypertension. In some embodiments, a subject having a low level ofsoluble ST2 compared to the reference level of soluble ST2 indicates hasa decreased risk (e.g., a risk decreased by at least 10%, 15%, 20%, 25%,30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or 80%) of developinghypertension.

Also provided are methods of evaluating the risk of developinghypertension in a subject that include determining a first level ofsoluble ST2 in a biological sample from a subject at a first time point,determining a second level of soluble ST2 in a biological sample from asubject at a second time point, comparing the second and first levels ofsoluble ST2, and identifying a subject having an elevated second levelof soluble ST2 compared to the first level of soluble ST2 as having anincreased risk of developing hypertension, or identifying a subjecthaving no significant change or a decreased second level of soluble ST2compared to the first level of soluble ST2 as having a decreased risk ofdeveloping hypertension. In some embodiments, a subject having anelevated second level of soluble ST2 (relative to the first level ofsoluble ST2) has an increased risk (e.g., an increased risk of at least10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%,80%, 85%, 90%, 95%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%,180%, 190%, or 200% higher than a reference subject, e.g., a controlsubject as described herein) of developing hypertension. In someembodiments, a subject having a low second level of soluble ST2 comparedto the first level of soluble ST2 indicates that the subject has adecreased risk (e.g., a risk decreased by at least 10%, 15%, 20%, 25%,30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or 80%) of developinghypertension.

The above methods may be used to determine the risk of developinghypertension within 3 years (e.g., risk of developing hypertensionwithin 3 years, within 1 year, within 9 months, within 6 months, orwithin 30 days of the time at which the biological sample was obtainedfrom the subject).

Some embodiments further include detecting a level of one or moreadditional biomarkers (e.g., any of the additional biomarkers describedherein, e.g., BNP, proBNP, and NT-proBNP) in a biological sample fromthe subject (e.g., the biological sample, the first biological sample,and/or the second biological sample). In these embodiments, a subjecthaving an elevation in the level of the one or more additionalbiomarkers in the biological sample as compared to a reference level ofthe one or more additional biomarkers is identified as having anelevated risk of developing hypertension, or a subject having nosignificant change or a decrease in the level of the one or moreadditional biomarkers as compared to the reference level of the one ormore additional biomarkers is identified as having a decreased risk ofdeveloping hypertension.

Some embodiments further include administering at least oneanti-hypertensive agent or performing increased cardiac monitoring on asubject identified as having an increased risk of developinghypertension (using any of the methods described herein), or performinglow frequency outpatient monitoring on a subject identified as having adecreased risk of developing hypertension (using any of the methodsdescribed herein).

Some embodiments include one or more of: updating a subject's clinicalfile (e.g., a computer readable medium) to indicate the subject'sdetermined risk of developing hypertension and suggested treatment basedon the subject's determined risk of developing hypertension, informing asubject identified as being at increased risk of developing hypertensionof symptoms of hypertension, instructing a subject identified as beingat increased risk of developing hypertension to self-monitor him orherself for symptoms of hypertension, instructing subject to implementchanges in lifestyle (e.g., perform or increase exercise therapy, changediet, reduce salt intake, and cease or decrease smoking), testing ordetermining the risk of hypertension in a lineal family member of asubject determined to have an increased risk of developing hypertension,performing one or more confirmatory diagnostic tests on a subjectidentified as having an increased risk of developing hypertension, andinforming a lineal family member about a subject's determined increasedrisk of developing hypertension.

Methods for Selecting a Subject for Participation in a Clinical Study

Also provided are methods of selecting a subject for participation in aclinical study of a treatment for reducing the risk of developinghypertension that include determining a level of soluble ST2 in abiological sample from a subject, comparing the level of soluble ST2 inthe biological sample to a reference level of soluble ST2, and selectinga subject having an elevated level of soluble ST2 in the biologicalsample compared to the reference level of soluble ST2 for participationin the clinical trial, or stratifying patients based on levels ofsoluble ST2. In some embodiments, a subject can be excluded fromparticipation in a clinical study of a treatment for reducing the riskof developing hypertension if the subject has a low level of soluble ST2in the biological sample compared to the reference level of soluble ST2(e.g., any of the reference levels of soluble ST2 described herein).

The clinical studies may be performed by a health care professional(e.g., a physician, a physician's assistant, a nurse, a phlebotomist, ora laboratory technician) in a health care facility (e.g., a hospital, aclinic, or a research center). The biological samples may be obtainedfrom subjects that present with one or more (e.g., at least two, three,four, or five) symptoms of a disease state (e.g., arrhythmia,cardiovascular disease, angina, or heart failure), subjects that areadmitted in a hospital, or subjects who are asymptomatic.

Some embodiments further include detecting a level of one or moreadditional biomarkers (e.g., any of the additional biomarkers describedherein, e.g., BNP, proBNP, and NT-proBNP) in a biological sample fromthe subject (e.g., the biological sample, the first biological sample,and/or the second biological sample). In these embodiments, a subjecthaving an elevation in the level of the one or more additionalbiomarkers in the biological sample as compared to a reference level ofthe one or more additional biomarkers is selected for participation in aclinical study of a treatment for reducing the risk of developinghypertension.

Additional factors may further indicate that the subject should beincluded in a clinical study of a treatment for reducing the risk ofdeveloping hypertension. Non-limiting examples of these additionalfactors include: prior diagnosis with cardiovascular disease, angina,heart attack, heart failure, renal failure, inflammation, or stroke; orpresentation of one or more (e.g., two, three, or four) of the followingsymptoms: shortness of breath, heart palpitations, increased heart rate,weakness, dizziness, nausea, sweating, chest discomfort or pressure,chest pain, arm pain, fullness, indigestion, sweating, wheezing, sleepapnea, and anxiety. Additional exemplary factors that indicate that asubject should be included in a clinical study of a treatment forreducing the risk of developing hypertension include a BMI of 25-30, aBMI of greater than 30, or continued therapy with one or more (e.g., atleast two, three, four, or five) pharmaceutical agents selected from thegroup of nitrates, calcium channel blockers, diuretics, thrombolyticagents, digitalis, renin-angiotensin-aldosterone system (RAAS)modulating agents (e.g., beta-adrenergic blocking agents,angiotensin-converting enzyme inhibitors, aldosterone antagonists, renininhibitors, and angiotensin II receptor blockers), andcholesterol-lowering agents (e.g., a statin).

Kits

Also provided are kits for use in a method described herein, containingan antibody that specifically binds to soluble ST2 and optionallyinstructions for using the kit (e.g., the antibodies in the kit) toperform any of the methods described herein. The antibody thatspecifically binds ST2 may be polyclonal, monoclonal, recombinant, e.g.,a chimeric or humanized, fully human, non-human, e.g., murine,mono-specific, or a single-chain antibody. Any of the kits describedherein may also be provided as an ELISA assay (e.g., may further includeone or more secondary antibodies and/or a substrate for detection). Forexample, any of the kits described herein may include an antibodyproduced from the hybridoma deposited at American Type CultureCollection and designated by Patent Deposit Designation PTA-10432, orany of the exemplary anti-ST2 antibodies described in WO 2011/127412 orU.S. Patent Application Publication No. 2011/0256635.

Any of the kits described herein may also include one or more (e.g.,two, three, four, or five) additional antibodies for one or more (e.g.,two, three, four, or five) additional markers selected from the groupof: proANP, NT-proANP, ANP, proBNP, NT-proBNP, BNP, troponin, CRP,galectin, creatinine, liver function enzymes, albumin, and bacterialendotoxin. Antibodies for ST2, galectin, proANP, NT-proANP, ANP, proBNP,NT-proBNP, BNP, troponin, CRP, creatinine, liver function enzymes,albumin, and bacterial endotoxin are commercially available.

The invention is further described in the following example, which doesnot limit the scope of the invention described in the claims.

EXAMPLE Example 1 Soluble ST2 can be Used to Assess the Risk ofDeveloping Hypertension in Apparently Healthy Subjects

A set of experiments was performed to determine if soluble ST2 (sST2) isuseful for predicting the occurrence of hypertension in apparentlyhealthy individuals. The FHS Offspring study is a prospective,observational, community-based study. Children (and spouses of thechildren) of FHS original-cohort participants were enrolled in 1971 andhave since been followed with serial examinations. Of 3,532 participantsattending the sixth examination (baseline, 1995-1998), 3,273participants returned for follow-up at the seventh examination(1998-2001). Of these, 52 were excluded due to missing BP data, 1,305were excluded due to prevalent hypertension at the baseline examination,34 due to missing sST2 measurement, 8 due to heart failure, 33 due tocoronary heart disease, 1 due to stage IV chronic kidney disease(defined as estimated glomerular filtration rate<30 ml/min/1.73 m²), and6 due to missing covariates, leaving 1,834 for inclusion in this study.All participants provided informed consent and the study was approved bythe appropriate Institutional Review Board.

In these experiments soluble ST2 was measured in stored samples from thesixth examination visit. The levels of soluble ST2 were determined usinga commercially available ELISA (Presage® ST2 Assay, Critical CareDiagnostics, San Diego, Calif.)(Dieplinger et a.l., Clin Chim Acta.2009; 409:33-40; Lu et al, Clin Chim Acta 2010; 411:1825-1826) accordingto the manufacturer's instructions.

The primary outcome to be assessed in this study was the time to thefirst occurrence of hypertension. Hypertension was defined as a systolicblood pressure≥140 mmHg, a diastolic blood pressure≥90 mmHg, or currentuse of antihypertensive medication. Change in systolic BP, diastolic BP,and pulse pressure was defined as the continuous change between thebaseline and follow-up examinations. Blood pressure was measured in theleft arm using a mercury sphygmomanometer, with the subject in a seatedposition after at least five minutes of rest. The examination BP was theaverage of two physician-obtained measurements.

Trends for increased risk of hypertension were found withlog-transformed (ln) sST2 (HR 1.21 [95% CI 1.06-1.39] in an age andgender adjusted model, P=0.006 and HR 1.22 [95% CI 1.05-1.42], in afully adjusted model, P=0.051 respectively). The full model includesage, gender, systolic and diastolic BP at baseline, diabetes mellitus,body-mass index, and smoking. The following Table 2 also shows that thehighest concentrations of sST2, as defined by the 4^(th) quartile,24.8-98.8 ng/ml, are also prognostic for hypertension in both theminimal and fully adjusted models.

TABLE 2 Age- and sex- Multivariable- adjusted model adjusted model* OR(95% CI) P OR (95% CI) P Log-sST2† 1.21 (1.06-1.39) 0.006 1.22(1.05-1.42) 0.01 Quartile 1 referent Referent Quartile 2 1.28(0.87-1.90) 1.34 (0.88-2.04) Quartile 3 1.52 (1.03-2.23) 1.43(0.94-2.18) Quartile 4 1.77 (1.20-2.62) 1.79 (1.17-2.73) P for trend0.003 0.008

In sum, the data show that measurement of sST2 can be used to assess therisk of developing hypertension in individuals who are otherwiseapparently healthy.

What is claimed is:
 1. A method of treating a healthy subject who does not have hypertension, the method comprising: a) determining a first level of soluble ST2 in a biological sample obtained from the subject at a first time point; (b) determining a second level of soluble ST2 in a biological sample obtained from the subject at a second time point, (c) selecting the subject for treatment with an anti-hypertensive drug when the subject has an elevated second level of soluble ST2 as compared to the first level of soluble ST2; and (d) administering the anti-hypertensive drug to the subject selected for treatment.
 2. The method of claim 1, wherein the biological sample in (a) and the biological sample in (b) comprise blood, serum, or plasma.
 3. The method of claim 1, further comprising: determining a first level of one of more additional biomarkers selected from the group consisting of: atrial natriuretic peptide (ANP), proANP, N-terminal (NT)-proANP, brain natriuretic peptide (BNP), proBNP, NT-proBNP, cardiac troponin I, C-reactive protein, creatinine, and Blood Urea Nitrogen (BUN) in a biological sample obtained from the subject at the first time point; determining a second level of the one or more additional biomarkers in a biological sample obtained from the subject at the second time point; and further selecting an anti-hypertensive drug for a subject having an elevated second level of the one or more additional biomarkers as compared to the first level of the one or more additional biomarkers.
 4. The method of claim 3, wherein the one or more additional biomarkers are selected from the group consisting of BNP, proBNP, and NT-proBNP.
 5. A method of treating a healthy subject who does not have hypertension, the method comprising: (a) determining a first level of soluble ST2 in a biological sample obtained from the subject at a first time point; (b) determining a second level of soluble ST2 in a biological sample obtained from the subject at a second time point; (c) identifying the subject as having an increased risk of developing hypertension when the subject has an elevated second level of soluble ST2 as compared to the first level of soluble ST2 ; and (d) administering an anti-hypertensive drug to the subject after identifying the subject as having an increased risk of developing hypertension.
 6. The method of claim 5, wherein the risk of developing hypertension is the risk of developing hypertension within three years.
 7. The method of claim 5, wherein the biological sample in (a) and the biological sample in (b) comprise blood, serum, or plasma.
 8. The method of claim 5, further comprising: determining a first level of one of more additional biomarkers selected from the group consisting of: atrial natriuretic peptide (ANP), proANP, N-terminal (NT)-proANP, brain natriuretic peptide (BNP), proBNP, NT-proBNP, cardiac troponin I, C-reactive protein, creatinine, and Blood Urea Nitrogen (BUN) in a biological sample obtained from the subject at the first time point; determining a second level of the one or more additional biomarkers in a biological sample obtained from the subject at the second time point; and further identifying a subject having an elevated second level of the one or more additional biomarkers as compared to the second level of the one or more additional biomarkers as having an increased risk of developing hypertension.
 9. The method of claim 8, wherein the one or more additional biomarkers are selected from the group consisting of BNP, proBNP, and NT-proBNP.
 10. The method of claim 1, wherein the subject is human.
 11. The method of claim 5, wherein the subject is human. 